using the RNA polymerase alpha subunit (rpoA) partial gene sequence specific primers, which also revealed the isolate as Lactobacillus plantarum (Genbank Accession Number KF286000).
The isolate DM5 showed broad antimicrobial activity as it inhibited the growth of Gram positive as well as Gram negative bacteria. The isolate DM5 had shown clear zone of inhibition against several major food borne pathogens, such as E. coli, S.
epidermis, S. aureus, A. feacalis and L. monocyotogenes in food stuffs, hence, it could be used as potential bio-preservative in foods. This is the first report of Lactobacillus plantarum showing antimicrobial activity as well as glucansucrase activity. The production of glucansucrase of isolate DM5 could be increased further by optimizing the fermentation process which could compensate the increasing demand of glucan as viscosifier and stabilizer in food industry. However, it is too early to claim the enzyme secreted by the isolate DM5 as glucansucrase. Further characterization of this enzyme is required to support this finding. This investigation also unravelled the abundance of industrially valuable microbial flora in fermented food products in North east India.
References
Aymerich, M.T., Garriga, M., Monfort, J.M., Nes, I. and Hugas, M. (2000) Bacteriocin-producing Lactobacilli in Spanish-style fermented sausages:
characterization of bacteriocins. Food Microbiol., 17, 33-45.
Barry, A.L. and Thornsberry, C. (1980) Susceptibility testing: diffusion procedures.
In: Manual of Clinical Microbiology (Lennette, E. H., Balows, A., Hausler, W.
J. Jr., Truant, J. P., ed). Washington, D.C: American Society for Microbiology, pp 463-74.
Bergey, D.H., John, G. H., Noel, R. K. And Peter, H.A.S. (1994) Bergey's Manual of Determinative Bacteriology (9th ed.). Lippincott Williams & Wilkins. ISBN 0- 683-00603-7.
Birnboim, H.C. and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res., 7, 1513-1523.
Bixquert, J.M. (2009) Treatment of irritable bowel syndrome with probiotics: an etiopathogenic approach at last. Rev. Esp. Enferm. Dig., 101, 553-564.
Briggs, M. (1953) The classification of lactobacilli by means of physiological tests. J.
Gen. Microbiol., 9, 234-248.
Buddhiman, T., Jyoti, P.T., Ulrich, S., Charles, M.A.P.F., Michael, G. and Wilhelm, H.H. (2008) Phenotypic and genotypic identification of lactic acid bacteria isolated from ethnic fermented bamboo tender shoots of North East India. Int.
J. Food Microbiol., 121, 35-40.
Cerning, J. (1990) Exocellular polysaccharides produced by lactic acid bacteria. FEMS Microbiol. Rev., 87, 113-130.
Corsetti, A., Lavermicocca, P., Morea, M., Baruzzi, F., Tosti, N. and Gobbetti M.
(2001) Phenotypic and molecular identification and clustering of lactic acid bacteria and yeasts from wheat (species Triticum durum and Triticum aestivum) sourdoughs of Southern Italy. Int. J. Food Microbiol., 64, 95-104.
Daming, R., Yinyu, W., Zilai, W., Jun, C., Hekui, L. and Jingye, Z. (2003) Complete DNA sequence and analysis of two cryptic plasmids isolated from Lactobacillus plantarum. Plasmid, 50, 70-73.
Danielsen, M. (2002) Characterization of the tetracycline resistance plasmid pMD5057 from Lactobacillus plantarum 5057 reveals a composite structure. Plasmid, 48, 98-103.
Danielsen, M. and Wind, A. (2003) Susceptibility of Lactobacillus spp. to antimicrobial agents. Int. J. Food Microbiol., 82, 1-11.
Das, D. and Goyal, A. (2010) Characterization and screening of antimicrobial activity of lactic acid bacterium isolated from a traditional beverage Marcha of Sikkim.
J Pharm. Chem., 4, 136-139.
Das, D. and Goyal, A. (2011) Expression of antagonism by lactic acid bacterium isolated from marcha: an ethnic fermented beverage of north-eastern Himalayas. Int. J. Biotechnol. Biochem., 7, 411-421.
De Man, J.C., Ragosa, M. and Sharpe, E. (1960) A medium for the cultivation of lactobacilli. J. Appl. Bact., 23, 130-135.
De, S., Kaur, G., Roy, A., Dogra, G., Kaushik, R., Yadav, P., Singh, R., Datta, T.K.
and Goswami, S.L. (2010) A Simple Method for the Efficient Isolation of Genomic DNA from Lactobacilli Isolated from Traditional Indian Fermented Milk (dahi). Indian J. Microbiol., 50, 412-418.
Dicks, L.M.T. and van Vuuren, H.I.I. (1987) A modification of the hot tube method for the detection of carbon dioxide produced by heterofermentative Lactobacillus strains. J. Microbiol. Meth., 6, 273-275.
Fugelsang, KC, and Edward CG (2007). Wine microbiology, Practical application and procedure. 2nd edition, Springer, 2, 29-44.
Galvez, A., Abriouel, H., Lopez, R.L. and Omar, N.B. (2007) Bacteriocin-based strategies for food biopreservation. Int. J. Food Microbiol., 120, 51–70.
Gram, C. (1884) Ueber die isolirte Farbung der Schizomyceten in Schnitt-und Trockenpraparaten. Fortschritte der Medcin., 2, 185-189.
Groenewald, W.H., van Reenen C.A., and Dicks, L.M.T. (2006) Strains of Lactobacillus plantarum in Grape Must are also Present in the Intestinal Tract of Vinegar Flies. S. Afr. J. Enol. Vitic., 27, 46-50.
Holzapfel, W.H., Geisen, R. and Schillinger, U. (1995) Biological preservation of foods with reference to protective cultures, bacteriocins and food-grade enzymes. Int. J. Food Microbiol., 24, 343–362.
Jeyaram, K., Singh, T.H., Romi, W., Devi, A.R., Singh, W.M., Dayanidhi, H., Singh, N.R. and Tamang, J.P. (2009) Traditional fermented foods of Manipur. Indian J. Traditional Knowl., 8, 115-121.
Jimenez-Diaz, R., Rios-Sanchez, R.M. and Piard, J.C. (1993) Plantaricins S and T, two new bacteriocins produced by Lactobacillus plantarum LPC010 isolated from a green olive fermentation. Appl. Environ. Microbiol., 59, 1416-1424.
Kanatani, K. and Oshimura, M. (1994) Plasmid-associated bacteriocin production by a Lactobacillus plantarum strain. Biosci. Biotechnol. Biochem., 58, 2084-2086.
Kandler, O. and Weiss, N. (1986) Genus Lactobacillus beijerinck 1901, 212AL,” In:
(Sneath, P. H. A. Mair, N. S. Sharpe, M. E. and Holt, J. G. ed.) Bergey’s manual of systematic bacteriology. Baltimore: Williams and Wilkins, 2, 209- 1234.
Kimura, M. (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J. Mol.
Evol., 16, 111-120.
Klare, I., Konstabe,l C., Werner, G., Huys, G., Vankerckhoven, V., Kahlmeter, G., Hildebrandt, B., Müller-Bertling, S., Witte, W. and Goossens, H. (2007) Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use. J.
Antimicrob. Chemother., 59, 900-912.
Kumar, K.S., Ganesan, K. and Subba-Rao, P.V. (2008) Antioxidant potential of solvent extracts of Kappaphycus alvarezii (Doty) Doty –An edible seaweed.
Food Chem., 107, 289-295.
Li, R., Zhai, Z., Yin, S., Huang, Y., Wang, Q., Luo, Y. and Hao, Y. (2009) Characterization of a Rolling-Circle Replication Plasmid pLR1 from Lactobacillus plantarum LR1. Curr. Microbiol., 58, 106-110.
Massi, M., Vitali, B., Federici, F., Matteuzzi, D. and Brigidi, P. (2004) Identification method based on PCR combined with automated ribotyping for tracking probiotic Lactobacillus strains colonizing the human gut and vagina. J. Appl.
Microbiol., 96, 777–786.
Messi, P., Bondi, M., Sabia, C., Battini, R. and Manicardi, G. (2001) Detection and preliminary characterization of a bacteriocin (plantaricin 35d) produced by a Lactobacillus plantarum strain. Int. J. Food Microbiol., 64, 193–198.
Naser, S.M., Dawyndt, P., Hoste, B., Gevers, D., Vandemeulebroecke, K., Cleenwerck, I., Vancanneyt, M. and Swings, J. (2007) Identification of lactobacilli by pheS and rpoA gene sequence analyses. Int. J. Syst. Evol.
Microbiol., 57, 2777–2789.
Nelson, N. (1944) A photometric adaptation of the Somoyogi method for the determination of glucose. J. Biol. Chem., 153, 375-380.
Nissen, L., Chingwaru, W., Sgorbati, B., Biavati, B. and Cencic A. (2009) Gut health promoting activity of new putative probiotic protective Lactobacillus spp.
Strains: a functional study in the small intestinal cell model. Int. J. Food Microbiol., 135, 288-294.
Patel, S. and Goyal, A. (2010) Isolation, characterization and mutagenesis of exopolysaccharide synthesizing new strains of lactic acid bacteria. Internet J.
Microbiol., 8, 1.
Purama, R.K., Agrawal, M., Majumder, A., Ahmed, S. and Goyal, A. (2008) Antibiotic sensitivity, carbohydrate fermentation characteristics and plasmid profiles of glucansucrase producing four Leuconostoc strains. J. Pure Appl.
Microbiol., 2, 139-146.
Purama, R.K., Goswami, P., Khan, A.T. and Goyal, A. (2009) Structural analysis and properties of dextran produced by Leuconostoc mesenteroides NRRL B-640.
Carbohydr. Polym., 76, 30-35.
Rai, AK., Palni, U. and Tamang, J.P. (2009) Traditional knowledge of the Himalayan people on production of indigenous meat products. Indian J. Traditional Knowl., 8, 104-109.
Ruiz-Barba, J.L., Piard, J.C. and Jimenez-Diaz, R. (1991) Plasmid profiles and curing of plasmids in Lactobacillus plantarum strains isolated from green olive fermentations. J. Appl. Bacteriol., 74, 417-421.
Saitou, N. and Nei, M. (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4, 406-425.
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, New York, Cold Spring Harbor Laboratory Press.
Satish-Kumar, R., Kanmani, P., Yuvraj, N., Paari, K.A., Pattukumar, V. and Arul V.
(2012) Traditional Indian fermented foods: a rich source of lactic acid bacteria.
Int. J. Food Sci. Nutr., 64, 1-14.
Shukla, S. and Goyal, A. (2011) 16S rRNA based identification of a glucan hyper- producing Weissella confusa. Enzym Res., 250842, doi:10.4061/2011/250842.
Singh, E.J., Singh, N.K.S. and Singh, N.R. (2009) Biodiversity conservation and natural resources in North East India with special reference to Manipur. NeBio 1, 42-47.
Somogyi, M. (1945) A new reagent for the determination of sugars. J. Biol. Chem., 160, 61-68.
Spelhaug, S.R. and Harlander, S.K. (1989) Inhibition of foodborne bacterial pathogens by bacteriocins from Lactobacillus lactis and Pediococcus pentosaceus. J.
Food Prot., 52, 856-862.
Swenson, J.M., Facklamand, R.R. and Thornsberry, C. (1990) Antimicrobial susceptibility of vancomycin-resistant Leuconostoc, Pediococcus, and Lactobacillus species. Antimicrob. Agents Chemother., 34, 543-549.
Tamang, J.P., Dewan, S., Tamang, B., Rai, A., Schillinger, U. and Holzapfel, W.H.
(2007) Lactic acid bacteria in Hamei and Marcha of North East India. Indian J. Microbiol., 47, 119-125.
Thapa, N., Pal, J. and Tamang, J.P. (2004) Microbial diversity in ngari, hentak and tungtap, fermented fish products of North-East India. World J. Microbiol.
Biotechnol., 20, 599-607.
Thokchom, S and Joshi, S.R. (2012) Antibiotic resistance and probiotic properties of dominant lactic microflora from Tungrymbai, an ethnic fermented soybean food of India. J. Microbiol., 50, 535-539.
Todorov, S.D. and Dicks, L.M.T. (2009) Effect of modified MRS medium on production and purification of antimicrobial peptide ST4SA produced by Enterococcus mundtii. Anaerobe., 15, 65-73.
Tsuchiya, H.M., Koepsell, H.J., Corman, J., Bryant, G., Bogard, M.O., Feger, V.H.
and Jackson, R.W. (1952) The effect of certain culture factors on production on dextransucrase by Leuconostoc mesenteroides. J. Bacteriol., 64, 521-526.
Ulrich, R.L. and Hughes, T.A. (2001) A rapid procedure for isolating chromosomal DNA from Lactobacillus species and other Gram-positive bacteria. Lett. Appl.
Microbiol., 32, 52-56.
Welman, A.D. and Maddox, I.S. (2003) Exopolysaccharides from lactic acid bacteria:
Perspectives and challenges. Trends Biotechnol., 21, 269–274.
Wu, C.W., Yin, L.J. and Jiang, S.T. (2004) Purification and Characterization of Bacteriocin from Pediococcus pentosaceus ACCEL. J. Agric. Food Chem., 52, 1146-1151.
Zeigler, D.R. (2003) Gene sequences useful for predicting relatedness of whole genomes in bacteria. Int. J. Syst. Evol. Microbiol., 53, 1893-1900.
Zhou, J.S., Pillidge, C.J., Gopal, P.K. and Gill, H.S. (2005) Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains. Int. J.
Food Microbiol., 98, 211-217.
Chapter 3
Probiotic attributes and antioxidative activity of Lactobacillus plantarum DM5